Autofluorescence imaging of A375 human melanoma cells: A pilot Quantitative In-Vitro Study
Afshan Shirkavand1,2, Ezeddin Mohajerani3, Shirin Farivar4, Leila Ataie-Fashtami5
1. Ph.D., Assistant prof of Biophotonics, Photodynamic Department, Medical Laser Research Center, Yara Institute, Tehran, Iran. Email: shirkavand@acecr.ac.ir
2. Laser and Plasma Research Institute, Shahid Beheshti University, Tehran, Iran
3. Ph.D., Professor of Photonics, Laser and Plasma Research Institute, Shahid Beheshti University, Tehran, Iran.
4. Ph.D., Associate Professor of Genetics, Stem Cells, Department of Cell and Molecular Biology, Faculty of Life Sciences and Biotechnology, Shahid Beheshti University, Tehran, Iran
5. M.D, Assistant Professor of Dermatology, Department of Regenerative Medicine, Royan Institute for Stem Cell Biology & Technology, ACECR, Tehran, Iran
Abstract
Background: Melanoma cancer is a common skin malignancy worldwide. There are several endogenous fluorophores in human skin which are potential for emitting inherent fluorescence, called intrinsic autofluorescence (AF). Melanin is known as an endogenous fluorophore in the epidermis basal cell layer. It seems to have approximately strong AF signal. This pilot study aimed to investigate the feasibility of the detection of AF signals in the A375 human melanoma cell line in the cell culture stage using an optical imaging system. Methods: The human skin melanoma cell line (A375) was cultured. For the imaging of the A375 human melanoma cell sample in this pilot study, the FluoVision optical imaging device (Tajhiz Afarinan Noori Parseh Co) was applied. An image processing MATLAB-based code was developed using the K-mean segmentation method. Results: A375 melanoma cell image fusion from raw data and AF processed images depicted based on the threshold AF signal higher than 40%. The percentage of the AF signal in A375 melanoma cells in our three cell samples was calculated as 3.11%±0.6. Conclusion: This imaging method has the advantage of no need for fluorophore labels over the existing fluorescence imaging methods, and it can be regarded as one of the important choices of Label-free imaging for this A375 melanoma cell line containing intrinsic endogenous fluorophore in cell studies.
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Speaker
Afshan Shirkavand
Assistant prof of Biophotonics, Photodynamic Department, Medical Laser Research Center, Yara Institute, Tehran, Iran
Iran
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